Mapping the murine cardiac 26S proteasome complexes

Aldrin V Gomes, Chenggong Zong, Ricky D. Edmondson, Xiaohai Li, Enrico Stefani, Jun Zhang, Richard C. Jones, Sheeno Thyparambil, Guang Wu Wang, Xin Qiao, Fawzia Bardag-Gorce, Peipei Ping

Research output: Contribution to journalArticle

135 Citations (Scopus)

Abstract

The importance of proteasomes in governing the intracellular protein degradation process has been increasingly recognized. Recent investigations indicate that proteasome complexes may exist in a species- and cell-type-specific fashion. To date, despite evidence linking impaired protein degradation to cardiac disease phenotypes, virtually nothing is known regarding the molecular composition, function, or regulation of cardiac proteasomes. We have taken a functional proteomic approach to characterize 26S proteasomes in the murine heart. Multidimensional chromatography was used to obtain highly purified and functionally viable cardiac 20S and 19S proteasome complexes, which were subjected to electrophoresis and tandem mass spectrometry analyses. Our data revealed complex molecular organization of cardiac 26S proteasomes, some of which are similar to what were reported in yeast, whereas others exhibit contrasting features that have not been previously identified in other species or cell types. At least 36 distinct subunits (17 of 20S and 19 of 19S) are coexpressed and assembled as 26S proteasomes in this vital cardiac organelle, whereas the expression of PA200 and 11S subunits were detected with limited participation in the 26S complexes. The 19S subunits included a new alternatively spliced isoform of Rpn10 (Rpn10b) along with its primary isoform (Rpn10a). Immunoblotting and immunocytochemistry verified the expression of key α and β subunits in cardiomyocytes. The expression of 14 constitutive α and β subunits in parallel with their three inducible subunits (β1i, β2i, and β5i) in the normal heart was not expected; these findings represent a distinct level of structural complexity of cardiac proteasomes, significantly different from that of yeast and human erythrocytes. Furthermore, liquid chromatography/tandem mass spectroscopy characterized 3 distinct types of post-translational modifications including (1) N-terminal acetylation of 19S subunits (Rpn1, Rpn5, Rpn6, Rpt3, and Rpt6) and 20S subunits (α2, α5, α7, β3, and β4); (2) N-terminal myristoylation of a 19S subunit (Rpt2); and (3) phosphorylation of 20S subunits (eg, α7)). Taken together, this report presents the first comprehensive characterization of cardiac 26S proteasomes, providing critical structural and proteomic information fundamental to our future understanding of this essential protein degradation system in the normal and diseased myocardium.

Original languageEnglish (US)
Pages (from-to)362-371
Number of pages10
JournalCirculation Research
Volume99
Issue number4
DOIs
StatePublished - Aug 2006

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Proteasome Endopeptidase Complex
Proteolysis
Proteomics
Protein Isoforms
Yeasts
Post Translational Protein Processing
Acetylation
Tandem Mass Spectrometry
Immunoblotting
Cardiac Myocytes
Liquid Chromatography
Organelles
Electrophoresis
Chromatography
Heart Diseases
Mass Spectrometry
Myocardium
Erythrocytes
Immunohistochemistry
Phosphorylation

Keywords

  • 19S and 20S proteasomes
  • Heart
  • Organelle proteomics
  • Protein degradation

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Gomes, A. V., Zong, C., Edmondson, R. D., Li, X., Stefani, E., Zhang, J., ... Ping, P. (2006). Mapping the murine cardiac 26S proteasome complexes. Circulation Research, 99(4), 362-371. https://doi.org/10.1161/01.RES.0000237386.98506.f7

Mapping the murine cardiac 26S proteasome complexes. / Gomes, Aldrin V; Zong, Chenggong; Edmondson, Ricky D.; Li, Xiaohai; Stefani, Enrico; Zhang, Jun; Jones, Richard C.; Thyparambil, Sheeno; Wang, Guang Wu; Qiao, Xin; Bardag-Gorce, Fawzia; Ping, Peipei.

In: Circulation Research, Vol. 99, No. 4, 08.2006, p. 362-371.

Research output: Contribution to journalArticle

Gomes, AV, Zong, C, Edmondson, RD, Li, X, Stefani, E, Zhang, J, Jones, RC, Thyparambil, S, Wang, GW, Qiao, X, Bardag-Gorce, F & Ping, P 2006, 'Mapping the murine cardiac 26S proteasome complexes', Circulation Research, vol. 99, no. 4, pp. 362-371. https://doi.org/10.1161/01.RES.0000237386.98506.f7
Gomes AV, Zong C, Edmondson RD, Li X, Stefani E, Zhang J et al. Mapping the murine cardiac 26S proteasome complexes. Circulation Research. 2006 Aug;99(4):362-371. https://doi.org/10.1161/01.RES.0000237386.98506.f7
Gomes, Aldrin V ; Zong, Chenggong ; Edmondson, Ricky D. ; Li, Xiaohai ; Stefani, Enrico ; Zhang, Jun ; Jones, Richard C. ; Thyparambil, Sheeno ; Wang, Guang Wu ; Qiao, Xin ; Bardag-Gorce, Fawzia ; Ping, Peipei. / Mapping the murine cardiac 26S proteasome complexes. In: Circulation Research. 2006 ; Vol. 99, No. 4. pp. 362-371.
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AU - Jones, Richard C.

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